Development of an in vitro co-culture system for elucidation of host-gut microbiota symbiotsis

• Project/Area Number: 18H02126
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Kyoto University
• Principal Investigator: Katayama Takane 京都大学, 生命科学研究科, 教授 (70346104)
• Co-Investigator(Kenkyū-buntansha): 加藤 紀彦 京都大学, 生命科学研究科, 助教 (40724612) ; 神戸 大朋 京都大学, 生命科学研究科, 准教授 (90303875)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2020: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2019: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2018: ¥10,530,000 (Direct Cost: ¥8,100,000、Indirect Cost: ¥2,430,000)
• Keywords: 共培養 / アピカル嫌気 / 嫌気性細菌 / 腸上皮細胞 / 腸管上皮細胞 / 偏性嫌気性細菌 / 腸上皮細胞株 / 共生 / 腸内細菌

Outline of Final Research Achievements

We have developed an apical anaerobic device in this study. The device was able to keep dissolved oxygen concentrations of apical side and basal side of Caco-2 monolayer at < 0.3 % and < 60 %, respectively, at least for 5 days. Transepithelial electrical resistance values were comparable to that of the cells maintained in CO2 incubator. Fluorescence immunostaining of claudin-2 revealed clear tight-junction formation of the cells. When obligate anaerobes belonging to Bacteroides and Bifidobacterium genera were added to the apical medium, the growth of these bacteria was stimulated by 10- to 1,000,000-folds. Transepithelial electrical resistance was comparable between before and after co-cultivation. These results indicate that the device is useful to examine cross-talk between intestinal epithelial cells and anaerobic microbes at the molecular levels.

Academic Significance and Societal Importance of the Research Achievements

本助成においてアピカル嫌気培養デバイスの有効性を示すことが出来た。本デバイスの特許出願に基づいた共同研究を国内企業2社と進めており、2022年もしくは2023年には実用化が見込まれている。本装置が汎用化されることで、よりin vivoに近い状態で共生・疾患メカニズムを解析することが可能となると考えられる。

Research Products

• [Int'l Joint Research] University College Cork(アイルランド)
• [Journal Article] Enzymatic Adaptation of Bifidobacterium bifidum to Host Glycans, Viewed from Glycoside Hydrolyases and Carbohydrate-Binding Modules (2020)
  • Author(s): Katoh Toshihiko、Ojima Miriam N.、Sakanaka Mikiyasu、Ashida Hisashi、Gotoh Aina、Katayama Takane
  • Journal Title: Microorganisms Volume: 8 Issue: 4 Pages: 481-481
  • DOI: 10.3390/microorganisms8040481
  • Peer Reviewed / Open Access
• [Journal Article] Evolutionary adaptation in fucosyllactose uptake systems supports bifidobacteria-infant symbiosis (2019)
  • Author(s): M. Sakanaka, M.E. Hansen, A. Gotoh, T. Katoh, K. Yoshida, T. Odamaki, H. Yachi, Y. Sugiyama, S. Kurihara, J. Hirose, T. Urashima, J. Xiao, M. Kitaoka, S. Fukiya, A. Yokota, L.L. Leggio, M.A. Hachem, T. Katayama
  • Journal Title: Science Advance Volume: 5 Issue: 8
  • DOI: 10.1126/sciadv.aaw7696
  • NAID: 120006716596
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] A plug flow apical anaerobic incubation device that enables co-culturing of anaerobic microbes and intestinal epithelial cells (2021)
  • Author(s): Aina GOTOH, Toyoyuki HASHIMOTO, Yasuko YONEDA, Yoichi FUJIYAMA, Eiichi OZEKI, Toshihiko KATOH, Yoh-ichi TAGAWA, Yun-Gi KIM, Koji HASE, Takane KATAYAMA
  • Organizer: 日本農芸化学会大会
• [Presentation] Symbiosis and co-evolution between bifidobacteria and humans, driven by human milk oligosaccharides (2019)
  • Author(s): Takane Katayama
  • Organizer: The 7th International conference on food factors
  • Int'l Joint Research / Invited
• [Presentation] 嫌気性腸内細菌と培養細胞の共培養システムの開発 (2018)
  • Author(s): 後藤愛那・永田千夏・加藤紀彦・神戸大朋・片山高嶺
  • Organizer: 第22回腸内細菌学会
• [Presentation] iGOEMONチャンバーの開発 (2018)
  • Author(s): 後藤愛那・神戸大朋・片山高嶺
  • Organizer: 2018年日本乳酸菌学会泊まり込みセミナー
• [Remarks]
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp/katayama/Member.html
• [Remarks]
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp/katayama/Home.html
• [Remarks]
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp/katayama/Home.html
• [Patent(Industrial Property Rights)] 嫌気性細菌などの細菌と上皮細胞との共培養装置及び共培養法 (2019)
  • Inventor(s): Takane Katayama, Taiho Kambe
  • Industrial Property Rights Holder: Takane Katayama, Taiho Kambe
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Overseas
• [Patent(Industrial Property Rights)] 嫌気性細菌などの細菌と上皮細胞との共培養装置及び共培養法 (2019)
  • Inventor(s): Takane Katayama, Taiho Kambe
  • Industrial Property Rights Holder: Takane Katayama, Taiho Kambe
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Overseas